mRNA decay is rapidly induced after spore germination of Saccharomyces cerevisiae
mRNA decay is rapidly induced after spore germination of Saccharomyces cerevisiae
复制标题
DOI：
10.1074/jbc.m206700200
复制
发表时间：
2002-10-25
影响因子：
4.8
通讯作者：
Lapeyre, B
中科院分区：
文献类型：
--
作者：
Brengues, M;Pintard, L;Lapeyre, B
当放置在有利的条件下时，来自酿酒酵母的孢子可以发芽并恢复营养生长。由于很难获得同步萌发的纯孢子群，因此对萌发的生化研究受到限制。在这里，我们报告可以通过离心淘洗根据孢子的大小来纯化和分选孢子，并且这些孢子能够同步发芽。同步它们的发育允许重新评估发芽的某些参数，并且我们证明在发芽诱导后转录和翻译都被非常迅速地诱导。孢子中含有的 mRNA 在 4 摄氏度下可稳定保存几个月。萌发诱导导致这些 mRNA 的快速降解，从而为研究真核生物中 mRNA 降解的诱导提供了一个简单的模型。来自孢子的 mRNA 被聚腺苷酸化、加帽，并与核糖体和多核糖体以及 mRNA 降解机制的组件一起沉积在蔗糖梯度上。孢子中多核糖体的存在使我们能够评估这些细胞中翻译装置的活性。我们提供的证据表明，在 30 摄氏度水中孵育的未发芽酵母孢子中存在持续的转录和翻译，这表明这些活动可能在孢子的长期存活中发挥作用。
Spores from the yeast Saccharomyces cerevisiae can germinate and resume their vegetative growth when placed in favorable conditions. Biochemical studies on germination have been limited by the difficulty of obtaining a pure population of spores germinating synchronously. Here, we report that spores can be purified and sorted according to their size by centrifugal elutriation and that these spores are able to germinate synchronously. Synchronizing their development has allowed reevaluating certain parameters of germination, and we demonstrate that both transcription and translation are induced very rapidly after germination induction. Spores contain mRNAs that are stable for several months in spores kept at 4 degreesC. Germination induction leads to very rapid degradation of these mRNAs, thus providing a simple model to study induction of mRNA decay in eukaryotes. mRNAs from the spore are polyadenylated, capped, and cosediment on sucrose gradients with ribosomes and polysomes and with components of the mRNA degradation machinery. The presence of polysomes in the spores led us to evaluate the activity of the translation apparatus in these cells. We present evidence that there is ongoing transcription and translation in nongerminating yeast spores incubated in water at 30 degreesC, suggesting that these activities could play a role in spore long term survival.